Caffeic acid protects against IL-1β-induced inflammatory responses and cartilage degradation in articular chondrocytes

doi:10.1016/j.biopha.2018.07.161

Biomedicine & Pharmacotherapy

Volume 107, November 2018, Pages 433-439

https://doi.org/10.1016/j.biopha.2018.07.161 Get rights and content

Highlights

•
CA down-regulates IL-1β-induced inflammatory responses.
•
CA protects chondrocytes from IL-1β-induced degradation of Collagen II and Aggrecan.
•
CA reduces the activation of NF-κB and JNK pathways induced by IL-1β.
•
CA may provide new avenues for osteoarthritis treatment.

Abstract

Osteoarthritis (OA) is a common articular disease that features cartilage loss and destruction. It has been confirmed that inflammation plays major roles in the progression of osteoarthritis. Caffeic acid (CA), a key dietary nutrient commonly found in coffee, has shown its anti-inflammatory properties in various inflammation diseases. However, the effects of CA in osteoarthritis remain explored. Here we investigated the effects of CA on IL-1β induced increased expression of inflammatory factors as well as the degradation of Collagen II and aggrecan in rat chondrocytes. CA prevented the cartilage damage induced by IL-1β in vivo organ culture of articular cartilage. Besides, the IL-1β induced increased production of inflammation factors such as iNOS and COX2 could be inhibited by CA. Additionally, CA could also suppress IL-1β induced expression of cartilage matrix catabolic enzymes such as ADAMTS5 and MMPs. Moreover, CA could prevent IL-1β induced degradation of Collagen II and aggrecan in chondrocytes. Furthermore, CA inhibited NF-κB activity and the activation of JNK pathway. This study reveals that CA inhibits IL-1β induced inflammation responses through suppression of NF-κB and MAPK related JNK signaling pathways. These results demonstrate that CA may provide new avenues for osteoarthritis treatment in future.

Graphical abstract

Introduction

Osteoarthritis (OA) is chronic degenerative joint disease in which patient experience clinical symptoms of joint pain and declining physical function [1]. It is a leading cause of disability among the elderly population and about 10% of men and 18% of women over 60 years old are affected by this disease [2]. Degeneration of articular cartilage is the most common cause of osteoarthritis. It is recognized as an irreversible process in the progression of OA [3]. There is no cure for osteoarthritis and treatment is limited to symptomatic relief or joint replacement [4]. Therefore, new treatment strategies are urgently need to delay the progression of OA.

An imbalance between anabolic and catabolic factors within chondrocytes can induce cartilage destruction. Patients with OA exhibit high levels of IL-1β in the joint cartilage as well as synovial tissues [5]. Work over the past year has reinforced the role of IL-1β in the onset and development of OA [6]. It is considered a key inflammatory cytokine involved in pathogenesis of OA. IL-1β has been shown to down-regulate the synthesis of cartilage matrix and inhibit the anabolic activity of chondrocytes [7]. Moreover, IL-1β directly induces the expression of genes encoding catabolic factors in chondrocytes, such as inducible Nitric Oxide Synthase (iNOS), Cyclooxygenase 2(COX2), aggrecanase-2(ADAMTS5) and matrix metalloproteinase (MMP1, MMP3 and MMP13) [8]. This dual effect makes IL-1β appear to be a reasonable target for OA treatment and medicines that can reverse the effects of IL-1β may provide a potential avenue for OA therapy.

Caffeic acid (3, 4-dihydroxy cinnamic acid, CA), a well-known phenolic phytochemical, is widely distributed in fruits, vegetables, wine and especially in coffee [9]. CA reportedly possess diverse biological potential including anti-oxidative [10], anti-inflammatory [11] and anticancer activities [12]. Previous studies indicated that CA was able to inhibit the gene expression of MMPs in lipopolysaccharide-activated human monocytes [13]. It also could inhibit solar ultraviolet radiation induced COX-2 expression in mouse skin [14]. Furthermore, another investigation indicated that intra-articular injections of caffeic acid phenethyl ester significantly decreased cartilage destruction and reduced loss of matrix proteoglycans in an experimental rabbit osteoarthritis model [15]. However, the exact role of CA in OA is still unclearly. In present study, we investigate the protective effects of CA in chondrocytes and its internal mechanism.

Section snippets

Antibodies and reagents

Recombinant rat IL-1β (501-RL-010)was obtained from R&D systems (Minneapolis, MN, USA). Antibody against MMP1 (Cat. No. 10371-2-AP) and P65 (Cat. No. 10745-1-AP) were obtained from Proteintech Group (Wuhan, China). Antibodies against aggrecan (Cat. No. ab36861) was purchased from Abcam (Cambridge, UK). Antibodies against Collagen II (Cat. No. sc-28887), iNOS (Cat. No. sc-7271) and MMP13 (Cat. No. sc-30073) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against

Effects of CA on chondrocyte viability

We first examined whether CA was toxic to chondrocytes. As shown in Fig. 1, the results of the CCK8 assay demonstrated that administration of CA (5, 10, or 20 μg/mL) had no significant effect on cell viability. Besides, we found that IL-1β treatment for 24 h couldn’t obviously decrease the cell viability.

CA suppressed IL-1β induced expression of iNOS and COX2 in chondrocytes

Western blot analysis was performed to detect the protein expression of the inflammation markers iNOS and COX2. Protein expression levels were normalized against GAPDH. As shown in Fig. 2,

Discussion

Osteoarthritis is the most common joint disease affecting large segments of the population. There is no radical therapy available for OA until now except for joint replacement. Therefore, research to find new natural products for the prevention and treatment of OA is gaining more and more attention [17]. Caffeic acid, a bioactive component of coffee, has possessed its ameliorative effect in capecitabineiInduced hepatic and renal dysfunction [18]. However, its protective effects on OA remain

Conclusions

This study reveals the potential ability of CA against osteoarthritis. It eﬀ ;ectively suppresses IL-1β induced degradation of collagen II and aggrecan and reduces the release of inﬂammatory mediators, including iNOS, COX2, MMPs and ADAMTS5. Furthermore, such suppressive eﬀ ;ects are likely to be carried out via NF-κB and MAPK related JNK pathway. Collectedly, our work provides evidence that CA may be developed as a medicine for the treatment of OA.

Author contributions

Hongbo You and Xiaojian Huang conceived the idea for the study. Xiaojian Huang carried out the research. Yang Xi, Qiyong Pan, Zekai Mao, Rui Zhang, Xiaohu MA analyzed and discussed the data. Hongbo You and Xiaojian Huang wrote the manuscript. All authors read and approved the ﬁnal manuscript.

Conﬂicts of interest

The authors declare no conﬂict of interest.

Acknowledgments

This research project was supported by the National Natural Science Foundation of China (Grant no. 81772390) and the Fundamental Research Funds for the Central Universities (grant no. 2017KFYXJJ104).

References (32)

A.E. Nelson
Osteoarthritis year in review 2017: clinical
Osteoarthr. Cartil.
(2018)
L. Troeberg et al.
Proteases involved in cartilage matrix degradation in osteoarthritis
Biochim. Biophys. Acta
(2012)
C. Manach et al.
Polyphenols: food sources and bioavailability
Am. J. Clin. Nutr.
(2004)
M.J. Chung et al.
Dietary phenolic antioxidants, caffeic acid and Trolox, protect rainbow trout gill cells from nitric oxide-induced apoptosis
Aquat. Toxicol.
(2006)
P. Vilela et al.
In vitro effect of caffeic acid phenethyl ester on matrix metalloproteinases (MMP-1 and MMP-9) and their inhibitor (TIMP-1) in lipopolysaccharide-activated human monocytes
Arch. Oral Biol.
(2015)
P. Chen et al.
The amelioration of cartilage degeneration by ADAMTS-5 inhibitor delivered in a hyaluronic acid hydrogel
Biomaterials
(2014)
A.D. Pearle et al.
Basic science of articular cartilage and osteoarthritis
Clin. Sports Med.
(2005)
B.C. Sondergaard et al.
MAPKs are essential upstream signaling pathways in proteolytic cartilage degradation--divergence in pathways leading to aggrecanase and MMP-mediated articular cartilage degradation
Osteoarthr. Cartil.
(2010)
S. Rigoglou et al.
The NF-kappaB signalling pathway in osteoarthritis
Int. J. Biochem. Cell Biol.
(2013)
J.S. Oh et al.
Biochanin-A antagonizes the interleukin-1beta-induced catabolic inflammation through the modulation of NFkappaB cellular signaling in primary rat chondrocytes
Biochem. Biophys. Res. Commun.
(2016)

M.B. Goldring et al.

Osteoarthritis

J. Cell. Physiol.

(2007)

F. Iannone et al.

The pathophysiology of osteoarthritis

Aging Clin. Exp. Res.

(2003)

P. Wojdasiewicz et al.

The role of inflammatory and anti-inflammatory cytokines in the pathogenesis of osteoarthritis

Mediators Inflamm.

(2014)

D.J. Blasioli et al.

The roles of catabolic factors in the development of osteoarthritis

Tissue Eng. Part B Rev.

(2014)

M.B. Goldring et al.

Transcriptional suppression by interleukin-1 and interferon-gamma of type II collagen gene expression in human chondrocytes

J. Cell. Biochem.

(1994)

S.Y. Sheu et al.

Arthropod steroid hormone (20-Hydroxyecdysone) suppresses IL-1beta-induced catabolic gene expression in cartilage

BMC Complement. Altern. Med.

(2015)

Cited by (41)

Caffeic acid modulates activation of neutrophils and attenuates sepsis-induced organ injury by inhibiting 5-LOX/LTB4 pathway
2023, International Immunopharmacology
Sepsis is a critical systemic inflammatory syndrome which usually leads to multiple organ dysfunction. Caffeic acid (CA), a phenolic compound derived from various plants, has been proved to be essential in neuroprotection, but its role in septic organ damage is unclear. This research aimed to investigate whether CA protects against organ injury in a mouse model of cecal ligation and puncture (CLP).
CA (30 mg/kg) or vehicle was administered by intraperitoneal injection immediately after CLP. The samples of blood, lungs, and livers were collected 24 h later. Organ injury was assessed by histopathological examination (HE staining), neutrophil infiltration (myeloperoxidase fluorescence), oxidative stress levels (MDA, SOD, HO-1), and inflammatory cytokines (TNF-α, IL-1β, and IL-6) release in lung and liver tissues. Neutrophil extracellular trap (NET) formation was analyzed by immunofluorescence. In vitro experiments were performed to investigate the potential mechanisms of CA using small interfering RNA (siRNA) techniques in neutrophils, and the effect of CA on neutrophil apoptosis was analyzed by flow cytometry.
Results showed that CA treatment improved the 7-day survival rate and attenuated the histopathological injury in the lung and liver of CLP mice. CA significantly reduced neutrophil infiltration in the lungs and livers of CLP mice. TNF-α, IL-1β, IL-6 and LTB4 were reduced in serum, lung, and liver of CA-treated CLP mice, and phosphorylation of MAPK (p38, ERK, JNK) and p65 NF-κB was inhibited in lungs and livers. CA treatment further increased HO-1 levels and enhanced superoxide dismutase (SOD) activity, but reduced malondialdehyde (MDA) levels and NET formation. Similarly, in vitro experiments showed that CA treatment and 5-LOX siRNA interference inhibited inflammatory activation and NET release in neutrophils, suppressed MAPK and NF-κB phosphorylation in LPS-treated neutrophils, and decreased LTB4 and cfDNA levels. Flow cytometric analysis revealed that CA treatment reversed LPS-mediated delayed apoptosis in human neutrophils, and Western blot also indicated that CA treatment inhibited Bcl-2 expression but increased Bax expression. CA treatment did not induce further changes in neutrophil apoptosis, inflammatory activation, and NET release when 5-LOX was knocked down by siRNA interference.
CA has a protective effect on lung and liver injury in a murine model of sepsis, which may be related to inhibition of the 5-LOX/LTB4 pathway.
Exploring active ingredients of anti-osteoarthritis in raw and wine-processed Dipsaci Radix based on spectrum-effect relationship combined with chemometrics
2023, Journal of Ethnopharmacology
Dipsaci Radix (DR) is the dry root of the Dipsacus asper Wall. ex DC., which has the function of tonifying the liver and kidney, continuing tendons and bones, and regulating blood vessels. However, there are few reports on the main active ingredients.
This study aimed to find the main active components of DR in the treatment of osteoarthritis (OA) by spectrum-effect relationship and compare the differences between RDR and WDR.
Firstly, the high-performance liquid chromatography (HPLC) method was used to establish the fingerprint of DR, and 10 peaks of them were determined by UPLC-Q-TOF/MS. Then, the OA rat model was established by injecting sodium iodoacetate to study the effect of DR on OA. The spectrum-effect relationship was analyzed by grey relational analysis (GRA) and Pearson correlation analysis.
According to the pharmacological results, compared with the model group, the cartilage score, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6), and Mankin score of rats in low, medium and high dose groups were decreased, and the therapeutic effect of wine-processed DR tended to be better than raw DR at the same dose. Finally, the active components of DR were preliminarily determined as 4 (loganic acid), 6 (chlorogenic acid), 8 (caffeic acid), 14 (dipsanoside B), 16, and 17 (asperosaponin VI) which had a large correlation in GRA and Pearson correlation analysis.
This study established the spectrum-effect relationship between the raw and wine-processed DR for the first time, which provided a theoretical basis for the study of the pharmacodynamic substance basis of DR before and after processing. This research provided a reference for the subsequent study of DR.
Potentials of natural antioxidants from plants as antiosteoporotic agents
2022, Studies in Natural Products Chemistry
Citation Excerpt :
Moreover, caffeic acid inhibited IL-1β–induced iNOS and COX2 production. These results provide interesting information that it may play a key role in preventing osteoporosis and osteoarthritis [80]. However, its low solubility has limited widespread application of caffeic acid in the biomedical field.
In osteoporosis, the density and quality of bone are reduced and become more fragile, with a significantly increased risk of fracture. Living tissue of bone undergoes continuous dynamic changes maintaining homeostasis by a delicate balance between osteoclasts and osteoblasts. Old bone is removed by osteoclasts (resorption), while new bone is added to the skeleton by osteoblasts (bone formation); an imbalance between these factors increases the risk of osteoporosis. Differentiation of osteoclasts and osteoblasts is strictly regulated by multiple cytokines, such as IL-1, IL-6, TNF-α, and TGF-β. Moreover, monocyte/macrophage colony stimulating factor and receptor activator of nuclear factor κB ligand induce osteoclast differentiation. In addition to these cytokines, it has become clear that reactive oxygen species regulate the differentiation of osteoclasts and osteoblasts.
Antioxidant phytochemicals like carotenoids, polyphenols, and organosulfur compounds have been found to reduce the risk of oxidative stress-related diseases, such as cancer, arteriosclerosis, and osteoporosis.
We focus on phytochemicals as antiosteoporotic agents in osteoporosis, and their antioxidative role via nuclear factor erythroid–derived related factor 2/Kelch-like ECH-associated protein pathway.
A novel caffeic acid derivative prevents renal remodeling after ischemia/reperfusion injury
2021, Biomedicine and Pharmacotherapy
Citation Excerpt :
It was discovered to contain abundant beneficial substances, including amino acids, polyphenols, flavonoids, and terpenes [25,27]. Caffeic acid and its derivatives are found in propolis and have high potential for preventing and treating diseases [28–30]. Caffeic acid phenethyl ester (CAPE), an important bioactive constituent of propolis, exhibits multiple benefits, such as antioxidant, anti-inflammatory, and anticancer properties [31–33].
Acute kidney disease due to renal ischemia/reperfusion (I/R) is a major clinical problem without effective therapies. The injured tubular epithelial cells may undergo epithelial–mesenchymal transition (EMT). It will loss epithelial phenotypes and express the mesenchymal characteristics. The formation of scar tissue in the interstitial space during renal remodeling is caused by the excessive accumulation of extracellular matrix components and induced fibrosis. This study investigated the effect of caffeic acid ethanolamide (CAEA), a novel caffeic acid derivative, on renal remodeling after injury. The inhibitory role of CAEA on EMT was determined by western blotting, real-time PCR, and immunohistochemistry staining. Treating renal epithelial cells with CAEA in TGF-β exposed cell culture successfully maintained the content of E-cadherin and inhibited the expression of mesenchymal marker, indicating that CAEA prevented renal epithelial cells undergo EMT after TGF-β exposure. Unilateral renal I/R were performed in mice to induce renal remodeling models. CAEA can protect against I/R-induced renal remodeling by inhibiting inflammatory reactions and consecutively inhibiting TGF-β-induced EMT, characterized by the preserved E-cadherin expression and alleviated α-SMA and collagen expression, as well as the alleviated of renal fibrosis. We also revealed that CAEA may exhibits biological activity by targeting TGFBRI. CAEA may antagonize TGF-β signaling by interacting with TGFBR1, thereby blocking binding between TGF-β and TGFBR1 and reducing downstream signaling, such as Smad3 phosphorylation. Our data support the administration of CAEA after I/R as a viable method for preventing the progression of acute renal injury to renal fibrosis.
Modulatory effects of caffeic acid on purinergic and cholinergic systems and oxi-inflammatory parameters of streptozotocin-induced diabetic rats
2021, Life Sciences
Citation Excerpt :
Nevertheless, our results showed that CA treatment was unable to reverse the increase in IL-1β expression in diabetic rats. By using a culture of rat chondrocytes, Huang et al. [58] demonstrated that CA treatment (20 μg/mL) negatively regulates the inflammatory responses induced by IL-1β. Thus, the results obtained here are probably because the anti-inflammatory effect of CA resides in the regulation of other proteins related to the inflammatory response and not the direct regulation of cytokines, as is the case of IL-1β and IL-6.
Diabetes mellitus (DM) is a metabolic disorder characterized by a chronic hyperglycemia state, increased oxidative stress parameters, and inflammatory processes.
To evaluate the effect of caffeic acid (CA) on ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase) and adenosine deaminase (ADA) enzymatic activity and expression of the A2A receptor of the purinergic system, acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) enzymatic activity and expression of the α7nAChR receptor of the cholinergic system as well as inflammatory and oxidative parameters in diabetic rats.
Diabetes was induced by a single dose intraperitoneally of streptozotocin (STZ, 55 mg/kg). Animals were divided into six groups (n = 10): control/oil; control/CA 10 mg/kg; control/CA 50 mg/kg; diabetic/oil; diabetic/CA 10 mg/kg; and diabetic/CA 50 mg/kg treated for thirty days by gavage.
CA treatment reduced ATP and ADP hydrolysis (lymphocytes) and ATP levels (serum), and reversed the increase in ADA and AChE (lymphocytes), BuChE (serum), and myeloperoxidase (MPO, plasma) activities in diabetic rats. CA treatment did not attenuate the increase in IL-1β and IL-6 gene expression (lymphocytes) in the diabetic state; however, it increased IL-10 and A2A gene expression, regardless of the animals' condition (healthy or diabetic), and α7nAChR gene expression. Additionally, CA attenuated the increase in oxidative stress markers and reversed the decrease in antioxidant parameters of diabetic animals.
Overall, our findings indicated that CA treatment positively modulated purinergic and cholinergic enzyme activities and receptor expression, and improved oxi-inflammatory parameters, thus suggesting that this phenolic acid could improve redox homeostasis dysregulation and purinergic and cholinergic signaling in the diabetic state.
Anti-arthritic and cartilage damage prevention via regulation of Nrf2/HO-1 signaling by glabridin on osteoarthritis
2021, Arabian Journal of Chemistry
Citation Excerpt :
The digested primary chondrocytes were cultured in DMEM supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin at 37 °C in a 5% CO2 incubator. Chondrocytes were seeded in a 96-well microtiter plate (2 × 105 cells/mL) and allowed for adhesion for 24 h following the method of Huang et al. (2018), using cell counting kit-8 (CCK-8) (Beyotime, Nanjing, China). Various concentrations (0, 10, 20, 40, 80, 100 µM) of glabridin (Sigma, St. Louis, USA) were applied on the chondrocytes and incubated for 72 h for determination of cytotoxicity.
Osteoarthritis is a common degenerative disease linked with inflammatory disorders and oxidative stress. Glabridin is an isoflavonoid and major active constituent of licorice. This study aims to investigate the anti-arthritic effects of glabridin on nuclear factor erythroid 2 – related factor 2 (Nrf2) signaling pathway, inflammatory responses, and cartilage degeneration in in-vitro and in-vivo models. Studies on IL-1β-induced chondrocyte model was performed to evaluate matrix metalloproteinase (MMP), Nrf2 signaling pathway. Glabridin was orally administered in monosodium-iodoacetate (MIA)-induced osteoarthritic (OA) rats for in-vivo evaluation. Pain and swelling of limbs were observed, oxidative stress markers and inflammatory cytokines were measured. Histomorphological changes in the joint cartilage were analyzed. Glabridin significantly reduced the arthritic score and paw swelling along with improved body weight, and organ index of rats. Histopathological score showed significant prevention of joint cartilage degeneration by glabridin. Expressions of TNF-α, IL-6, IL-10, and IL-1β were attenuated by glabridin (p < 0.05) in MIA-induced rats. Protein expressions of iNOS, COX-2, ADAMTS5, MMP-3, and MMP-13 were suppressed (p < 0.05) whereas the Nrf2/HO-1 signaling was activated by glabridin (p < 0.05) in osteoarthritic chondrocytes. Therefore, anti-arthritic and chondroprotective activity of glabridin is suggested by the inhibition of MMP expression and regulation of Nrf2/HO-1 signaling.

View all citing articles on Scopus

View full text